Barley, a fundamental cereal grain, is extensively grown for its nutritional advantages and adaptability in food and beverage manufacturing. Hence, this study aimed at profiling the fatty acid composition and quantifying the crude fat content and the individual fatty acid constituents in barley samples taken from three sub-districts of Dangla district. the barley samples were collected from Debaca, Chara, and Sahara sub-district and the crude fat and fatty acid constituents were determined following a standard procedure. Results revieweld that the crude fat contet of three samples ranged from 4.85-5.95% by mass. In addition, atotal of twelve fatty acids, Seven saturated and five unsaturated fatty acids were detected in all barley samples. The highest fatty acid content of barley in three sub-district were myristic, Linolenic ,Palmitic, oleic ,linoleic, Arachidic, gondoic, behenic, Palmitoleic, Margaric ,stearic, and lignoceric acid. Palmitic acid was the most abundant fatty acid in the barley and accounted for 44.2–49.9% of the total fatty acid content. The level of Linoleic acid found in this study ranged from28.47% - 32.22% of the total fatty acid. Furthermore, the total content of saturated fatty acids fell between 266 and 311 mg/100g, at its pure content of 210-266 mg/100 g, with palmitic acid being the most predominant contributor. Myristic and stearic acids were other members of saturated fatty acids contributing to its total value. On the other hand, unsaturated fatty acids are between 166- 227mg/100g in terms of total concentration, with linoleic acid, between 121-169mg/100g, being the biggest contributor, and included in this class are the dominating Oleic and gondoic acids, ranging from 44-60 mg/100g and 2-4 mg/100 respectively. It could, therefore, be inferred from the data presented that the saturated fats were higher in number compared to their unsaturated counterparts in the barley samples. There is no significant difference in the mean cocnetration of the fatty acid between the sampling sub districts. Keywords: Barley; Fatty acids; Gas Chromatography Mass Spectrometry